HI,
Many of us who use New Objective emitters are experiencing similar supply
issues. I have used their emitters exclusively for a number of years.
My last order was this past spring. I am down to my last 20 emitters- and
have no idea what to do when I run out. We have run 1000's of patient samples
with this set up on multiple MS insturments.
After countless time of trying to reach New Objective- I finally have a ship
date of October 20th. We have not been able to shut down due to COVID, and our
MS core needs to continue to run.
So I feel Turck's pain.
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 9:45 AM, "ABRF Discussion Forum on behalf of turck"
<email obscured> on behalf of <email obscured>> wrote:
EXTERNAL MESSAGE
_________________________________________________________________
Dear MassSpecers, we have been using PicoFrit Self-Pack Column: 360
μm OD 75 (article no: PF360-75-10-N-5) that we pack ourselves. Our last
order for the columns was placed in March and we still have not received them.
Can anybody recommend a similar product from a different vendor that is
available? Thanks much! Chris
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If you have the budget, the Pharmafluidics columns may be an option?
My alternative is to not use the picofrit columns.
Instead I have in the past made a blunt ended column plugged with a self-made
sol-gel frit and mate that to a stainless steel PepSep emitter and take a bit
of a hit on peak resolution. I prefer to use a Sutter laser puller, then pull
tips and pack them. It is important that the packing goes all the way to the
very tip of column with no dead space between the packed bed and the tip; and
to run a standard digest such as a HeLa, yeast or E. coli to QC each column.
If either of these steps fail then pack another one. I have found that a
column with a small gap from the end of the packed bed to the tip results in
inferior chromatographic peak shape and clogs more quickly compared to a column
with the packed bed all the way to the tip which is more robust and long lived.
At first, it may (will) take some time and a few (or a lot of) failures to pull
a good tip and also pack a good column but with concerted patient effort it
becomes fairly routine. I favor a Sutter laser program which produces a
fairly short tip that after separation has a very fine point, too fine, and
then I very very gently with almost no pressure drag it briefly across 2000
grit lapping paper to knock off the really fine part of the tip. This results
in a reproducible tip opening ~10µ which packs very nicely ... most of the time
... as long as the bed doesn't "keystone" before hitting the tip opening. I
use 80% isopropanol:20% acetonitrile as the packing solvent and use an Accela
600 pump to pack at ~500bar. I don't have any success using Helium and a
stirred cell for packing. Those I've packed with gas seem to unpack as the
pressure comes to atmosphere no matter how long I let it depressurize.
Yes, it's tedious and failure rate can be high until you get the hang of it. I
would like to find a way to fabricate a frit into a pulled tip, but so far
haven't had success with that.
Best,
Brian
Brian Hampton
Proteomics Core Lab
Center for Vascular and Inflammatory Diseases
University of Maryland School of Medicine
BioPark One, Rm 301
800 West Baltimore Street
Baltimore, MD 21201
410-706-8207
On 7/21/20, 9:57 AM, "ABRF Discussion Forum on behalf of Robert.Salzler"
<email obscured> on behalf of <email obscured>> wrote:
CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS email
system. Whether the sender is known or not known, hover over any links before
clicking and use caution opening attachments.
HI,
Many of us who use New Objective emitters are experiencing similar supply
issues. I have used their emitters exclusively for a number of years.
My last order was this past spring. I am down to my last 20 emitters- and
have no idea what to do when I run out. We have run 1000's of patient samples
with this set up on multiple MS insturments.
After countless time of trying to reach New Objective- I finally have a
ship date of October 20th. We have not been able to shut down due to COVID,
and our MS core needs to continue to run.
So I feel Turck's pain.
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 9:45 AM, "ABRF Discussion Forum on behalf of turck"
<email obscured> on behalf of <email obscured>> wrote:
EXTERNAL MESSAGE
_________________________________________________________________
Dear MassSpecers, we have been using PicoFrit Self-Pack Column: 360
μm OD 75 (article no: PF360-75-10-N-5) that we pack ourselves. Our last
order for the columns was placed in March and we still have not received them.
Can anybody recommend a similar product from a different vendor that is
available? Thanks much! Chris
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Hi Brian,
some comments on pressure packing using Helium. Depending on the C18 material,
different solvents behave different, some make a real fine slurry with free
C18-particles, others create bigger aggregates of the C18 material, what is not
good at all. I watched through a binocular. As far as I remember, my favorite
Reprosil was best with Methanol. To pack the column finally, I used an old
outdated ABI 120B (?) double syringe pump, run it at quite high flow rate until
overpressure, then stopped it and left the column overnight just releasing the
pressure. These columns had been very stable and reliable.
I even took my pressure devise to Cuba to train students there, had a lot of
fun at the airport during the control.
We used the BSA digest (now from BRUKER) to test our columns, 100 fmole
injections.
Manfred
Am 21.07.20, 16:42 schrieb "ABRF Discussion Forum im Auftrag von Hampton,
Brian" <email obscured> im Auftrag von <email obscured>>:
If you have the budget, the Pharmafluidics columns may be an option?
My alternative is to not use the picofrit columns.
Instead I have in the past made a blunt ended column plugged with a
self-made sol-gel frit and mate that to a stainless steel PepSep emitter and
take a bit of a hit on peak resolution. I prefer to use a Sutter laser puller,
then pull tips and pack them. It is important that the packing goes all the
way to the very tip of column with no dead space between the packed bed and the
tip; and to run a standard digest such as a HeLa, yeast or E. coli to QC each
column. If either of these steps fail then pack another one. I have found
that a column with a small gap from the end of the packed bed to the tip
results in inferior chromatographic peak shape and clogs more quickly compared
to a column with the packed bed all the way to the tip which is more robust and
long lived.
At first, it may (will) take some time and a few (or a lot of) failures to
pull a good tip and also pack a good column but with concerted patient effort
it becomes fairly routine. I favor a Sutter laser program which produces a
fairly short tip that after separation has a very fine point, too fine, and
then I very very gently with almost no pressure drag it briefly across 2000
grit lapping paper to knock off the really fine part of the tip. This results
in a reproducible tip opening ~10µ which packs very nicely ... most of the time
... as long as the bed doesn't "keystone" before hitting the tip opening. I
use 80% isopropanol:20% acetonitrile as the packing solvent and use an Accela
600 pump to pack at ~500bar. I don't have any success using Helium and a
stirred cell for packing. Those I've packed with gas seem to unpack as the
pressure comes to atmosphere no matter how long I let it depressurize.
Yes, it's tedious and failure rate can be high until you get the hang of
it. I would like to find a way to fabricate a frit into a pulled tip, but so
far haven't had success with that.
Best,
Brian
Brian Hampton
Proteomics Core Lab
Center for Vascular and Inflammatory Diseases
University of Maryland School of Medicine
BioPark One, Rm 301
800 West Baltimore Street
Baltimore, MD 21201
410-706-8207
On 7/21/20, 9:57 AM, "ABRF Discussion Forum on behalf of Robert.Salzler"
<email obscured> on behalf of <email obscured>> wrote:
CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS
email system. Whether the sender is known or not known, hover over any links
before clicking and use caution opening attachments.
HI,
Many of us who use New Objective emitters are experiencing similar
supply issues. I have used their emitters exclusively for a number of years.
My last order was this past spring. I am down to my last 20 emitters-
and have no idea what to do when I run out. We have run 1000's of patient
samples with this set up on multiple MS insturments.
After countless time of trying to reach New Objective- I finally have
a ship date of October 20th. We have not been able to shut down due to COVID,
and our MS core needs to continue to run.
So I feel Turck's pain.
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 9:45 AM, "ABRF Discussion Forum on behalf of turck"
<email obscured> on behalf of <email obscured>> wrote:
EXTERNAL MESSAGE
_________________________________________________________________
Dear MassSpecers, we have been using PicoFrit Self-Pack Column: 360
μm OD 75 (article no: PF360-75-10-N-5) that we pack ourselves. Our
last order for the columns was placed in March and we still have not received
them. Can anybody recommend a similar product from a different vendor that is
available? Thanks much! Chris
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Does anyone happen to know what New Objective uses for their distal coating?
http://www.newobjective.com/products/emitters/off_vs_on.shtml
Thanks
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 10:42 AM, "ABRF Discussion Forum on behalf of Hampton, Brian"
<email obscured> on behalf of <email obscured>> wrote:
If you have the budget, the Pharmafluidics columns may be an option?
My alternative is to not use the picofrit columns.
Instead I have in the past made a blunt ended column plugged with a
self-made sol-gel frit and mate that to a stainless steel PepSep emitter and
take a bit of a hit on peak resolution. I prefer to use a Sutter laser puller,
then pull tips and pack them. It is important that the packing goes all the
way to the very tip of column with no dead space between the packed bed and the
tip; and to run a standard digest such as a HeLa, yeast or E. coli to QC each
column. If either of these steps fail then pack another one. I have found
that a column with a small gap from the end of the packed bed to the tip
results in inferior chromatographic peak shape and clogs more quickly compared
to a column with the packed bed all the way to the tip which is more robust and
long lived.
At first, it may (will) take some time and a few (or a lot of) failures to
pull a good tip and also pack a good column but with concerted patient effort
it becomes fairly routine. I favor a Sutter laser program which produces a
fairly short tip that after separation has a very fine point, too fine, and
then I very very gently with almost no pressure drag it briefly across 2000
grit lapping paper to knock off the really fine part of the tip. This results
in a reproducible tip opening ~10µ which packs very nicely ... most of the time
... as long as the bed doesn't "keystone" before hitting the tip opening. I
use 80% isopropanol:20% acetonitrile as the packing solvent and use an Accela
600 pump to pack at ~500bar. I don't have any success using Helium and a
stirred cell for packing. Those I've packed with gas seem to unpack as the
pressure comes to atmosphere no matter how long I let it depressurize.
Yes, it's tedious and failure rate can be high until you get the hang of
it. I would like to find a way to fabricate a frit into a pulled tip, but so
far haven't had success with that.
Best,
Brian
Brian Hampton
Proteomics Core Lab
Center for Vascular and Inflammatory Diseases
University of Maryland School of Medicine
BioPark One, Rm 301
800 West Baltimore Street
Baltimore, MD 21201
410-706-8207
On 7/21/20, 9:57 AM, "ABRF Discussion Forum on behalf of Robert.Salzler"
<email obscured> on behalf of <email obscured>> wrote:
CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS
email system. Whether the sender is known or not known, hover over any links
before clicking and use caution opening attachments.
HI,
Many of us who use New Objective emitters are experiencing similar
supply issues. I have used their emitters exclusively for a number of years.
My last order was this past spring. I am down to my last 20 emitters-
and have no idea what to do when I run out. We have run 1000's of patient
samples with this set up on multiple MS insturments.
After countless time of trying to reach New Objective- I finally have
a ship date of October 20th. We have not been able to shut down due to COVID,
and our MS core needs to continue to run.
So I feel Turck's pain.
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 9:45 AM, "ABRF Discussion Forum on behalf of turck"
<email obscured> on behalf of <email obscured>> wrote:
EXTERNAL MESSAGE
_________________________________________________________________
Dear MassSpecers, we have been using PicoFrit Self-Pack Column: 360
μm OD 75 (article no: PF360-75-10-N-5) that we pack ourselves. Our
last order for the columns was placed in March and we still have not received
them. Can anybody recommend a similar product from a different vendor that is
available? Thanks much! Chris
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I think it is a metal that is sputtered onto the tip in a manner such as used
in the electron microscopy field. In the paper noted below they say they
obtained gold coated tips from New Objective. If you really need to apply
voltage at the tip using a distal coating, then you might try doing this
yourself using polyaniline (PANI) as done here:
https://www.sciencedirect.com/science/article/pii/S1044030500001343
"Polyaniline: a conductive polymer coating for durable nanospray emitters"
Their composition used for coating was: "... xylene-soluble conductive PANI
dispersions (Mw 117,000 Da, 48.16% xylenes, 12.62% butyl cellosolve, the
remainder solid PANI)"
Sigma sells a product similar to what was used in that paper: product no.
576379 Polyaniline (emeraldine base) average Mw ~100,000
I have not tried this, just providing info for alternatives given the situation
with New Objective.
I instead use an electrical junction at the head of the column with a small SS
union from Valco on this page: https://www.vici.com/cfit/360um_met.php
These small fittings feature single use connection SS nuts which I do not use.
Instead I use the PEEK nuts shown on this page:
https://www.vici.com/cfit/360um_pk.php instead of the SS nuts that come with
the SS union. They have remained leak free at least to 980 bar in my
experience. You have to be very careful during assembly to prevent breaking
the fused silica in the SS union. I use an insulated wire connected to the
voltage source and an alligator clip on the SS union to deliver the HV. Low
tech, but functional. I prefer to use the 150u bore union for longer life
before clogging.
You can also make your own conductive TEE out of the Valco nano all PEEK
fittings by inserting a ~360u diameter gold wire into one of the ports of the
TEE. This makes for a very light weight HV junction but provides a much
smaller surface area for delivering the HV to the mobile phase compared to the
union. Not sure if this makes any difference. A platinum wire might be better
than gold? I noticed that the end of the gold wire in contact with the mobile
phase becomes discolored in a short time. Maybe the gold wire I'm getting is
low quality. I thought gold was inert.
Best regards,
Brian
On 7/23/20, 5:04 PM, "ABRF Discussion Forum on behalf of Robert.Salzler"
<email obscured> on behalf of <email obscured>> wrote:
Does anyone happen to know what New Objective uses for their distal
coating?
http://www.newobjective.com/products/emitters/off_vs_on.shtml
Thanks
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 10:42 AM, "ABRF Discussion Forum on behalf of Hampton, Brian"
<email obscured> on behalf of <email obscured>> wrote:
If you have the budget, the Pharmafluidics columns may be an option?
My alternative is to not use the picofrit columns.
Instead I have in the past made a blunt ended column plugged with a
self-made sol-gel frit and mate that to a stainless steel PepSep emitter and
take a bit of a hit on peak resolution. I prefer to use a Sutter laser puller,
then pull tips and pack them. It is important that the packing goes all the
way to the very tip of column with no dead space between the packed bed and the
tip; and to run a standard digest such as a HeLa, yeast or E. coli to QC each
column. If either of these steps fail then pack another one. I have found
that a column with a small gap from the end of the packed bed to the tip
results in inferior chromatographic peak shape and clogs more quickly compared
to a column with the packed bed all the way to the tip which is more robust and
long lived.
At first, it may (will) take some time and a few (or a lot of) failures
to pull a good tip and also pack a good column but with concerted patient
effort it becomes fairly routine. I favor a Sutter laser program which
produces a fairly short tip that after separation has a very fine point, too
fine, and then I very very gently with almost no pressure drag it briefly
across 2000 grit lapping paper to knock off the really fine part of the tip.
This results in a reproducible tip opening ~10µ which packs very nicely ...
most of the time ... as long as the bed doesn't "keystone" before hitting the
tip opening. I use 80% isopropanol:20% acetonitrile as the packing solvent and
use an Accela 600 pump to pack at ~500bar. I don't have any success using
Helium and a stirred cell for packing. Those I've packed with gas seem to
unpack as the pressure comes to atmosphere no matter how long I let it
depressurize.
Yes, it's tedious and failure rate can be high until you get the hang
of it. I would like to find a way to fabricate a frit into a pulled tip, but
so far haven't had success with that.
Best,
Brian
Brian Hampton
Proteomics Core Lab
Center for Vascular and Inflammatory Diseases
University of Maryland School of Medicine
BioPark One, Rm 301
800 West Baltimore Street
Baltimore, MD 21201
410-706-8207
On 7/21/20, 9:57 AM, "ABRF Discussion Forum on behalf of
Robert.Salzler" <email obscured> on behalf of
<email obscured>> wrote:
CAUTION: This message originated from a non UMB, UMSOM, FPI, or
UMMS email system. Whether the sender is known or not known, hover over any
links before clicking and use caution opening attachments.
HI,
Many of us who use New Objective emitters are experiencing similar
supply issues. I have used their emitters exclusively for a number of years.
My last order was this past spring. I am down to my last 20
emitters- and have no idea what to do when I run out. We have run 1000's of
patient samples with this set up on multiple MS insturments.
After countless time of trying to reach New Objective- I finally
have a ship date of October 20th. We have not been able to shut down due to
COVID, and our MS core needs to continue to run.
So I feel Turck's pain.
Robert R. Salzler
Regeneron Pharmaceuticals, Inc.
Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162
777 Old Saw Mill River Road
Tarrytown, NY 10591-06707
Office: 914-847-3075
Cell: 716-864-6787
On 7/21/20, 9:45 AM, "ABRF Discussion Forum on behalf of turck"
<email obscured> on behalf of <email obscured>> wrote:
EXTERNAL MESSAGE